Hydrolytic properties of a β-mannosidase purified from Aspergillus niger

Pia Ademark, J. Lundqvist, Maija Tenkanen, N. Torto, Henrik Stålbrand (Corresponding Author), F. Tjerneld, Per Hägglund

Research output: Contribution to journalArticleScientificpeer-review

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Abstract

A β-mannosidase was purified to homogeneity from the culture filtrate of Aspergillus niger. A specific activity of 500 nkat mg−1 and a 53-fold purification was achieved using ammonium sulfate precipitation, anion-exchange chromatography, and gel filtration. The isolated enzyme has an isoelectric point of 5.0 and appears to be a dimer composed of two 135-kDa subunits. It is a glycoprotein and contains 17% N-linked carbohydrate by weight. Maximal activity was observed at pH 2.4–5.0 and at 70°C. The β-mannosidase hydrolyzed β-1,4-linked manno-oligosaccharides of degree of polymerization (DP) 2–6 and also released mannose from polymeric ivory nut mannan and galactomannan. The Km and Vmax values for p-nitrophenyl-β-d-mannopyranoside were 0.30 mM and 500 nkat mg−1, respectively. Hydrolysis of d-galactose substituted manno-oligosaccharides showed that the β-mannosidase was able to cleave up to, but not beyond, a side group. An internal peptide sequence of 15 amino acids was highly similar to that of an Aspergillus aculeatus β-mannosidase belonging to family 2 of glycosyl hydrolases.
Original languageEnglish
Pages (from-to)281-289
Number of pages9
JournalJournal of Biotechnology
Volume75
Issue number2-3
DOIs
Publication statusPublished - 1999
MoE publication typeA1 Journal article-refereed

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Mannosidases
Oligosaccharides
Aspergillus niger
Aspergillus
Hydrolases
Glycoproteins
Carbohydrates
Mannose
Chromatography
Dimers
Peptides
Purification
Amino acids
Hydrolysis
Ion exchange
Negative ions
Gels
Enzymes
Polymerization
Mannans

Cite this

Ademark, P., Lundqvist, J., Tenkanen, M., Torto, N., Stålbrand, H., Tjerneld, F., & Hägglund, P. (1999). Hydrolytic properties of a β-mannosidase purified from Aspergillus niger. Journal of Biotechnology, 75(2-3), 281-289. https://doi.org/10.1016/S0168-1656(99)00172-8
Ademark, Pia ; Lundqvist, J. ; Tenkanen, Maija ; Torto, N. ; Stålbrand, Henrik ; Tjerneld, F. ; Hägglund, Per . / Hydrolytic properties of a β-mannosidase purified from Aspergillus niger. In: Journal of Biotechnology. 1999 ; Vol. 75, No. 2-3. pp. 281-289.
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abstract = "A β-mannosidase was purified to homogeneity from the culture filtrate of Aspergillus niger. A specific activity of 500 nkat mg−1 and a 53-fold purification was achieved using ammonium sulfate precipitation, anion-exchange chromatography, and gel filtration. The isolated enzyme has an isoelectric point of 5.0 and appears to be a dimer composed of two 135-kDa subunits. It is a glycoprotein and contains 17{\%} N-linked carbohydrate by weight. Maximal activity was observed at pH 2.4–5.0 and at 70°C. The β-mannosidase hydrolyzed β-1,4-linked manno-oligosaccharides of degree of polymerization (DP) 2–6 and also released mannose from polymeric ivory nut mannan and galactomannan. The Km and Vmax values for p-nitrophenyl-β-d-mannopyranoside were 0.30 mM and 500 nkat mg−1, respectively. Hydrolysis of d-galactose substituted manno-oligosaccharides showed that the β-mannosidase was able to cleave up to, but not beyond, a side group. An internal peptide sequence of 15 amino acids was highly similar to that of an Aspergillus aculeatus β-mannosidase belonging to family 2 of glycosyl hydrolases.",
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Ademark, P, Lundqvist, J, Tenkanen, M, Torto, N, Stålbrand, H, Tjerneld, F & Hägglund, P 1999, 'Hydrolytic properties of a β-mannosidase purified from Aspergillus niger', Journal of Biotechnology, vol. 75, no. 2-3, pp. 281-289. https://doi.org/10.1016/S0168-1656(99)00172-8

Hydrolytic properties of a β-mannosidase purified from Aspergillus niger. / Ademark, Pia; Lundqvist, J.; Tenkanen, Maija; Torto, N.; Stålbrand, Henrik (Corresponding Author); Tjerneld, F.; Hägglund, Per .

In: Journal of Biotechnology, Vol. 75, No. 2-3, 1999, p. 281-289.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Ademark, Pia

AU - Lundqvist, J.

AU - Tenkanen, Maija

AU - Torto, N.

AU - Stålbrand, Henrik

AU - Tjerneld, F.

AU - Hägglund, Per

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AB - A β-mannosidase was purified to homogeneity from the culture filtrate of Aspergillus niger. A specific activity of 500 nkat mg−1 and a 53-fold purification was achieved using ammonium sulfate precipitation, anion-exchange chromatography, and gel filtration. The isolated enzyme has an isoelectric point of 5.0 and appears to be a dimer composed of two 135-kDa subunits. It is a glycoprotein and contains 17% N-linked carbohydrate by weight. Maximal activity was observed at pH 2.4–5.0 and at 70°C. The β-mannosidase hydrolyzed β-1,4-linked manno-oligosaccharides of degree of polymerization (DP) 2–6 and also released mannose from polymeric ivory nut mannan and galactomannan. The Km and Vmax values for p-nitrophenyl-β-d-mannopyranoside were 0.30 mM and 500 nkat mg−1, respectively. Hydrolysis of d-galactose substituted manno-oligosaccharides showed that the β-mannosidase was able to cleave up to, but not beyond, a side group. An internal peptide sequence of 15 amino acids was highly similar to that of an Aspergillus aculeatus β-mannosidase belonging to family 2 of glycosyl hydrolases.

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M3 - Article

VL - 75

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JO - Journal of Biotechnology

JF - Journal of Biotechnology

SN - 0168-1656

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Ademark P, Lundqvist J, Tenkanen M, Torto N, Stålbrand H, Tjerneld F et al. Hydrolytic properties of a β-mannosidase purified from Aspergillus niger. Journal of Biotechnology. 1999;75(2-3):281-289. https://doi.org/10.1016/S0168-1656(99)00172-8